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Neural/vascular network based on animal adipose tissue-derived hydrogel and construction method and application thereof

A neural network and animal fat technology, applied in the field of biomedicine, can solve problems such as limited capacity, large differences, complicated and lengthy induction process, etc., to achieve a good three-dimensional surface structure and avoid immune rejection.

Pending Publication Date: 2022-04-29
SOUTH UNIVERSITY OF SCIENCE AND TECHNOLOGY OF CHINA
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] However, there are certain defects in related technologies. The neural tissue formed by directional induction of stem cells is complicated and lengthy on the one hand, and the structure of the neural network formed on the other hand is relatively simple. Other methods are needed to reproduce the complex neural network.
The three-dimensional neural network constructed solely by microfluidic technology only provides a scaffold for the construction of the neural network, but cannot completely copy the microenvironment of neural tissue growth.
However, the three-dimensional neural network constructed by synthetic polymer hydrogel usually contains relatively simple biological components and cannot provide nutrients for the neural network due to the large difference between the mechanical stiffness of the polymer hydrogel and the brain tissue environment. Limited ability to mimic the biological composition of native tissue

Method used

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  • Neural/vascular network based on animal adipose tissue-derived hydrogel and construction method and application thereof
  • Neural/vascular network based on animal adipose tissue-derived hydrogel and construction method and application thereof
  • Neural/vascular network based on animal adipose tissue-derived hydrogel and construction method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0108] Example 1: Porcine adipose tissue decellularization treatment

[0109] 1. Obtaining Porcine Adipose Acellular Matrix

[0110] Pig adipose tissue was extracted under aseptic conditions, the blood clot was washed with deionized water, the adipose tissue was cut into small pieces of 1 mm × 1 mm and rinsed with deionized water for 2 to 3 times, and the chopped adipose tissue was placed in Store at -80°C and 37°C alternately and repeat freeze-thaw 3 times.

[0111]Add 2 volumes of deionized water to the frozen-thawed adipose tissue and homogenize for 10 minutes, centrifuge at 4°C and 1200rpm for 10 minutes, after centrifugation, the adipose tissue is separated into layers, take the bottom white precipitate, add 1% Triton Shake the -X100 at 25°C for 1h, then wash with deionized water for 30min, repeat 3 times and centrifuge at 4°C, 1200rpm for 10min, collect the precipitate and add 100% isopropanol to shake overnight, centrifuge to remove residual lipid quality ingredients....

Embodiment 2

[0116] Example 2: Acquisition and cultivation of neuronal cells and vascular cells

[0117] 1. Extraction and culture of neurons

[0118] Decapitated SD suckling mice (purchased from Guangdong Medical Animal Center) 1-2 days old on ice, and peeled off their brains, put them in 4°C pre-cooled PBS, cut the corpus callosum with ophthalmic forceps under a dissecting microscope, The two hemispheres of the brain were spread out to both sides, and the cortex tissue and hippocampal tissue were peeled off respectively with ophthalmic forceps, and placed in 4°C pre-cooled serum-free DMEM / F12 medium (Gibco, product number: C11330500BT). Cut the tissue into 1mm×1mm small pieces and transfer them to 15mL centrifuge tubes, add 1mL of 2.5% trypsin solution, and digest at 37°C for 15min. After digestion, add F12 medium to stop digestion and gently rinse three times to remove trypsin solution. Blow and beat the tissue 5-10 times with a sterile pipette, put the cell suspension into a centrifu...

Embodiment 3

[0121] Example 3: Construction of a three-dimensional neuronal network based on porcine adipose tissue-derived hydrogel

[0122] 1. Preparation of Porcine Adipose Tissue-Derived Hydrogels

[0123] At 4°C, take the acellular matrix solution A in Example 1, add 0.1 mol / L NaOH solution to adjust the pH to 7, and use 10×PBS to adjust the osmotic pressure to obtain the acellular matrix solution B. Add the acellular matrix solution B into the square microfluidic chip, and form a hydrogel at 37°C for about 30 minutes.

[0124] 2. Scanning electron microscope observation of porcine adipose tissue-derived hydrogel

[0125] Using scanning electron microscopy to observe the surface microstructure of porcine adipocyte extracellular matrix hydrogel, the results are as follows figure 2 shown. in figure 2 The B is figure 2 A Magnification of the white dotted box, from figure 2 It can be seen from the results that the porcine adipose acellular matrix hydrogel prepared by the present...

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Abstract

The invention relates to a nerve / blood vessel network based on animal adipose tissue-derived hydrogel and a construction method and application thereof, the nerve / blood vessel network comprises a three-dimensional neural network and an artificial blood vessel, the three-dimensional neural network uses hydrogel prepared from an acellular matrix of animal adipose tissue as a substrate to culture nerve cells, the neural network with at least two layers of cell structures is constructed in a stacking manner, and the preparation material of the hydrogel can be taken from autologous adipose tissue, so that the neural / vascular network based on the animal adipose tissue source hydrogel constructed by the invention can be transplanted to the autologous body and is free from immunological rejection. The neural / vascular network based on the animal adipose tissue-derived hydrogel can accurately simulate the microenvironment of neural network growth, can monitor the electrical signal change of neuron activity in real time, and is of great significance in promoting brain science research.

Description

technical field [0001] The invention belongs to the field of biomedicine, and in particular relates to a nerve / vascular network based on hydrogel derived from animal adipose tissue and its construction method and application. Background technique [0002] In related technologies, although there has been some progress in the related research on the construction of a three-dimensional neuron network, due to the complexity of the brain environment, people's research on the growth and development of nerves in the brain and signal transduction is not very clear. In order to intuitively monitor the activity of neurons in the brain, it is of great significance to construct a regular neural network in vitro and make it perform normal signal transduction, regulatory feedback and other functions. Using electrical stimulation to induce the growth of neural networks, simulating the three-dimensional neural network of the autologous brain and monitoring changes in electrical signals of n...

Claims

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Application Information

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IPC IPC(8): C12N5/0793C12N5/071G01R19/00G01N27/00C12Q1/02
CPCC12N5/0619C12N5/0691G01R19/0084G01N27/00G01N33/5082C12N2513/00C12N2533/90
Inventor 蒋兴宇刘晓艳
Owner SOUTH UNIVERSITY OF SCIENCE AND TECHNOLOGY OF CHINA
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